Light-regulated phosphorylation of maize phosphoenolpyruvate carboxykinase plays a vital role in its activity

Chao, Qing; Liu, Xiaoyu; Mei, Ying‐Chang; Gao, Zili; Chen, Yi‐Bo; Qian, Chunrong; Hao, Yunhong; Wang, Baichen

doi:10.1007/s11103-014-0171-3

Cited by 22 publications

(28 citation statements)

References 44 publications

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“…In addition, C 4 plants possess another abundant phosphate driven transporter, the phosphoenolpyruvate phosphate translocator (PPT), which is essential for the transport of PEP from the chloroplast in MC (Brautigam et al, 2008;Majeran et al, 2008). Also, the activities of the critical enzymes involved in C 4 carbon assimilation, such as PEPC, PCK, and pyruvate phosphate dikinase, are modulated by reversible phosphorylation (Ashton and Hatch, 1983;Jiao and Chollet, 1991;Chao et al, 2014).…”

Section: Phosphorusmentioning

confidence: 99%

Ensuring Nutritious Food Under Elevated CO2 Conditions: A Case for Improved C4 Crops

2020

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Global climate change is a challenge for efforts to ensure food security for future generations. It will affect crop yields through changes in temperature and precipitation, as well as the nutritional quality of crops. Increased atmospheric CO 2 leads to a penalty in the content of proteins and micronutrients in most staple crops, with the possible exception of C 4 crops. It is essential to understand the control of nutrient homeostasis to mitigate this penalty. However, despite the importance of mineral nutrition for plant performance, comparably less is known about the regulation of nutrient uptake and homeostasis in C 4 plants than in C 3 plants and mineral nutrition has not been a strong focus of the C 4 research. Here we review what is known about C 4 specific features of nitrogen and sulfur assimilation as well as of homeostasis of other essential elements. We identify the major knowledge gaps and urgent questions for future research. We argue that adaptations in mineral nutrition were an integral part of the evolution of C 4 photosynthesis and should be considered in the attempts to engineer C 4 photosynthetic mechanisms into C 3 crops.

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Section: Phosphorusmentioning

confidence: 99%

Ensuring Nutritious Food Under Elevated CO2 Conditions: A Case for Improved C4 Crops

2020

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“…Malate activates AthPEPCK1 in the direction of PEP synthesis (Table 2) and, at the same time, it inhibits PEPC, which uses PEP [58][59][60][61][62][63][64], providing the necessary regulation ( Figure 5A). As previously mentioned, plant PEPCKs are phosphorylated [29,46,[65][66][67][68][69]. Leegood and Walker [67] proposed that this mechanism might coordinate the activities of both enzymes.…”

Section: Discussionmentioning

confidence: 90%

Biochemical characterization of phosphoenolpyruvate carboxykinases from Arabidopsis thaliana

Rojas

Hartman

Figueroa

et al. 2019

Biochemical Journal

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ATP-dependent phosphoenolpyruvate carboxykinases (PEPCKs, EC 4.1.1.49) from C4 and CAM plants have been widely studied due to their crucial role in photosynthetic CO2 fixation. However, our knowledge on the structural, kinetic and regulatory properties of the enzymes from C3 species is still limited. In this work, we report the recombinant production and biochemical characterization of two PEPCKs identified in Arabidopsis thaliana: AthPEPCK1 and AthPEPCK2. We found that both enzymes exhibited high affinity for oxaloacetate and ATP, reinforcing their role as decarboxylases. We employed a high-throughput screening for putative allosteric regulators using differential scanning fluorometry and confirmed their effect on enzyme activity by performing enzyme kinetics. AthPEPCK1 and AthPEPCK2 are allosterically modulated by key intermediates of plant metabolism, namely succinate, fumarate, citrate and α-ketoglutarate. Interestingly, malate activated and glucose 6-phosphate inhibited AthPEPCK1 but had no effect on AthPEPCK2. Overall, our results demonstrate that the enzymes involved in the critical metabolic node constituted by phosphoenolpyruvate are targets of fine allosteric regulation.

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“…Despite the fact that there are no phosphorylation sites described for PEPCK or RuBisCO large subunit in the UniProt database, some recent publications reported several phosphorylation sites. For PEPCK, four phosphorylated residues (S55, T58, T59, and T120) were reported in 2014 [ 18 ], and an additional one (S119) was published in 2015 [ 14 ]. For RuBisCO large subunit, Bi et al .…”

Section: Resultsmentioning

confidence: 99%

Selection of an Appropriate Protein Extraction Method to Study the Phosphoproteome of Maize Photosynthetic Tissue

et al. 2016

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Often plant tissues are recalcitrant and, due to that, methods relying on protein precipitation, such as TCA/acetone precipitation and phenol extraction, are usually the methods of choice for protein extraction in plant proteomic studies. However, the addition of precipitation steps to protein extraction methods may negatively impact protein recovery, due to problems associated with protein re-solubilization. Moreover, we show that when working with non-recalcitrant plant tissues, such as young maize leaves, protein extraction methods with precipitation steps compromise the maintenance of some labile post-translational modifications (PTMs), such as phosphorylation. Therefore, a critical issue when studying PTMs in plant proteins is to ensure that the protein extraction method is the most appropriate, both at qualitative and quantitative levels. In this work, we compared five methods for protein extraction of the C4-photosynthesis related proteins, in the tip of fully expanded third-leaves. These included: TCA/Acetone Precipitation; Phenol Extraction; TCA/Acetone Precipitation followed by Phenol Extraction; direct extraction in Lysis Buffer (a urea-based buffer); and direct extraction in Lysis Buffer followed by Cleanup with a commercial kit. Protein extraction in Lysis Buffer performed better in comparison to the other methods. It gave one of the highest protein yields, good coverage of the extracted proteome and phosphoproteome, high reproducibility, and little protein degradation. This was also the easiest and fastest method, warranting minimal sample handling. We also show that this method is adequate for the successful extraction of key enzymes of the C4-photosynthetic metabolism, such as PEPC, PPDK, PEPCK, and NADP-ME. This was confirmed by MALDI-TOF/TOF MS analysis of excised spots of 2DE analyses of the extracted protein pools. Staining for phosphorylated proteins in 2DE revealed the presence of several phosphorylated isoforms of PEPC, PPDK, and PEPCK.

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Light-regulated phosphorylation of maize phosphoenolpyruvate carboxykinase plays a vital role in its activity

Cited by 22 publications

References 44 publications

Ensuring Nutritious Food Under Elevated CO2 Conditions: A Case for Improved C4 Crops

Ensuring Nutritious Food Under Elevated CO2 Conditions: A Case for Improved C4 Crops

Biochemical characterization of phosphoenolpyruvate carboxykinases from Arabidopsis thaliana

Selection of an Appropriate Protein Extraction Method to Study the Phosphoproteome of Maize Photosynthetic Tissue

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